脂质体转染反义脱氧寡核苷酸对K562细胞α珠蛋白基因表达及细胞增殖的影响  

Effect of Liposomal Transfection of Antisense Oligodeoxynucleotide on Alpha-globin Gene Expression and Proliferation of K562 Cells

作　　者：刘容容[1] 赖永榕[1] 马劼[1] 

机构地区：[1]广西医科大学第一附属医院血液内科,南宁530021

出　　处：《中国实验血液学杂志》2007年第5期1065-1069,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目;编号30360034

摘　　要：本研究查明脂质体转染反义脱氧寡核苷酸(ASON)对K562细胞α珠蛋白基因表达的抑制作用及对细胞增殖的影响,探讨基因调控治疗β地中海贫血的新思路。设计合成靶向α珠蛋白基因的ASON,并与正义寡核苷酸(SON)和空白对照进行比较。采用脂质体转染技术将ASON、SON与K562细胞共同培养,用荧光显微术、流式细胞术检测脂质体转染效率,用实时荧光定量RT-PCR法检测K562细胞中α珠蛋白基因表达情况,并通过Cell Counting Kit-8(CCK-8)法观察脂质体转染ASON技术对细胞增殖的影响。结果表明:脂质体转染效率在ASON作用24h达到最高,脂质体转染ASON组的α珠蛋白基因表达水平显著低于SON和空白对照组(p<0.01),并对细胞的增殖有明显的抑制作用,上述效应呈浓度依赖性。结论:脂质体转染ASON能特异性的抑制K562细胞中α珠蛋白基因表达,可能会成为β地中海贫血基因治疗的一个新靶点。The objective of study was to investigate the effect of liposomal transfection of antisense oligodeoxynucleotide （ASON） on alpha-globin gene expression and proliferation of K562 cells, to explore the new way of gene therapy in β-thalassemia. Targeted ASON of alpha-globin was designed and synthesized, and compared with positive control [ sense oligodeoxynucleotide （SON） group] and blank control, By liposomal transfection, ASON, SON was co-cultured with K562. The efficiency of trasfection was assayed by fluorescence microscopy and flow cytometry （FCM）, the alpha-globin gene expression of K562 was measured by real-time PCR, and the proliferation of K562 was determined by Cell Count Kit-8 assay. The results indicated that the highest efficiency was at 24 hours after liposomal transfection, the gene expression level of alpha-globin in ASON group was significantly lower than that in SON group and blank control （p 〈 0.01 ）. The proliferation of K562 cells was obviously inhibited, meanwhile the above effect showed the dose-dependent manner. It is concluded that the liposomal transfection of ASON inhibits the alpha-globin gene expression of K562 cells, which may be the new target for gene therapy in β-thalassemia.

关 键 词：Α珠蛋白基因 脂质体转染 反义脱氧寡核苷酸 K562细胞 细胞增殖 地中海贫血 

分 类 号：R556.61[医药卫生—血液循环系统疾病]